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Construction of a bovine bacterial artificial chromosome library.

Abstract

We have constructed a bacterial artificial chromosome (BAC) library for bovine use in genome mapping. Currently, the library consists of 14,000 clones which achieves a 50% probability (P=0.5) of the library containing a specific unique DNA sequence. Sixty thousand clones or about three haploid bovine genomes will be required to achieve a 95 /o probability (P=0.95) of containing a unique sequence. An average insert size of 142kb was estimated from the analysis of 68 randomly selected BAC clones. Polymerase chain reaction (PCR) was used to screen the exist bovine BAC library. For three microsatellite markers and three anchor locus markers, positive clones were found for one microsatellite marker (ETH225) and two anchor locus markers (Glucocerebrocidase and 3(J-hydroxy-5-ene steroid dehydrogenase). Characterization of the BAC library, in terms of the percentage of chimeric clones and the stability of inserts in BAC clones is in progress.